Mass spectrometry-based proteomics is an emerging technology that has seen possible applications everywhere. The technique can potentially use one drop of our blood to reveal onset and progress of diseases. Current R&D using plasma/serum proteome sample sets, however, is severely limited by the high complexity and dynamic range of protein concentrations. Vast majority of plasma/serum proteomic strategies lack adequate reproducibility, sensitivity, and robustness for clinical tests. Solutions for these serious technological barriers to develop a reliable detection platform toward clinical implementation are elusive. In this NIH STTR Phase I study, we will develop a novel analytical platform based on functionalized soluble nanopolymers into commercial products for simple, routine serum/plasma biomarker discovery. Through this STTR Phase I project, we will leap from our expertise in pioneering the application of soluble nanopolymers to establish Polymer-based Metal Ion Affinity Capture (PolyMAC) functionalized with different metal ions as a powerful platform for sequential depletion of high-abundant proteins and isolation of low abundant plasma proteins to achieve sensitive, reproducible and quantitative plasma proteomics. In particular, we will validate the platform through analyzing plasma samples including those from Ossabaw swine, a distinctive large animal model for early detection of coronary artery disease (CAD). In order to achieve these goals, the following aims will be completed: Aim #1: Optimization of PolyMAC-Cu for high-abundant plasma protein depletion. Aim #2: Optimization of in-depth plasma proteome analyses using sequential PolyMAC-M isolations. By the completion of STTR Phase I studies, we expect to develop a versatile sample preparation platform with core reagents and kits for plasma proteomics and to facilitate translating proteomics discovery to clinical applications.